Ndjomou Jean, Liu Ying, O'Malley John, Ericsson Maria, He Johnny J
Department of Microbiology and Immunology, Indiana University School of Medicine, R2 302, 950 W. Walnut Street, Indianapolis, IN 46202, USA.
Biochem Biophys Res Commun. 2007 Jul 20;359(1):57-62. doi: 10.1016/j.bbrc.2007.05.073. Epub 2007 May 21.
Invention of subgenomic HCV replicon a few years ago and recent success of in vitro production of infectious HCV have improved our knowledge of the HCV life cycle, replication, pathogenesis, and screening of anti-HCV therapeutics. However, the highly genotype-dependent nature of the in vitro HCV production system has limited its potential for HCV research. In this study, we constructed a recombinant DNA-based HCV system that contained EF-1alpha promoter-driven HCV genotype 1b with HCV E1/E2 deleted and replaced by GFP. We co-transfected this recombinant cDNA with HCV E1/E2 or VSV-G expression plasmid into 293T cells, and we showed HCV protein expression and processing and demonstrated production of HCV-like particles in culture supernatant of co-transfected cells. These results support potential use of this system for studies on expression and processing of the HCV polyprotein and assembly and release of HCV-like particles.
几年前亚基因组丙型肝炎病毒（HCV）复制子的发明以及近期传染性HCV体外生产的成功，提高了我们对HCV生命周期、复制、发病机制以及抗HCV治疗药物筛选的认识。然而，体外HCV生产系统高度依赖基因型的特性限制了其在HCV研究中的潜力。在本研究中，我们构建了一种基于重组DNA的HCV系统，该系统包含由EF-1α启动子驱动的缺失HCV E1/E2并被绿色荧光蛋白（GFP）取代的HCV 1b基因型。我们将该重组cDNA与HCV E1/E2或水疱性口炎病毒糖蛋白（VSV-G）表达质粒共转染到293T细胞中，并且我们展示了HCV蛋白的表达和加工过程，并证明在共转染细胞的培养上清液中产生了HCV样颗粒。这些结果支持该系统在HCV多蛋白表达和加工以及HCV样颗粒组装和释放研究中的潜在应用。
